基于冷冻肿瘤组织切片的磷酸化HER2免疫组织化学染色方法探讨

Exploration of Phospho-HER2 Immunohistochemistry Staining on Frozen Tumor Tissues

目的探索针对磷酸化人类表皮生长因子受体2(human epidermal growth factor receptor 2,HER2)免疫组织化学染色实验中冷冻组织切片的最佳固定与保存条件。方法在BALB/c-nu/nu小鼠中建立人源性乳腺癌细胞BT474的皮下移植瘤模型,对比不同的抗原固定条件和存储时间下,免疫组织化学染色观察肿瘤模型中磷酸化HER2的信号变化。结果冰冻切片用体积分数为10%的甲醛溶液固定5 min,可以维持良好的细胞形态。在-20℃的存储条件下,肿瘤组织中磷酸化HER2蛋白在12周内表达稳定。结论冰冻组织切片进行免疫组织化学染色用时短,可以作为常规石蜡切片染色方法的补充或者替代。

Objective To investigate the optimized fixation and storage conditions for the immunohistochemistry detection of phospho-human epidermal growth factor receptor 2(HER2)on frozen sections.Methods Xenograft model was established by subcutaneously injecting human breast cancer cell BT474 into the abdomen of BALB/c-nu/nu mice.The mice were sacrificed when the tumor volume up to 300 mm^3.The tumor samples were quickly frozen,sectioned,fixed in different reagents and stored for different time period.Phospho-HER2 immunohistochemistry was performed to evaluate the change of protein expression under these conditions.Results Good cell morphology was kept by the fixation of frozen section in 50% formalin for 5 minutes.The expression of phosphoHER2 protein was stable within 12 weeks after fixation when the tumor tissues were stored in-20℃.Conclusion The immunohistochemistry staining on frozen tissues takes less time and can be a supplement or replacement of traditional staining on formalin-fixed and paraffin-embedded sections.